A new promoterless reporter vector reveals antisense transcription in Mycoplasma genitalium
A new promoterless reporter vector reveals antisense transcription in Mycoplasma genitalium
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DOI：
10.1099/mic.0.2006/007559-0
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发表时间：
2007-08-01
期刊：
影响因子：
2.8
通讯作者：
Pinol, Jaurne
中科院分区：
文献类型：
--
作者：
Lluch-Senar, Maria;Vallmitjana, Miquel;Pinol, Jaurne
The mechanisms that promote and regulate transcription in mycoplasmas are poorly understood. Here, a promoter-probe vector based on the pMTnTetM438 minitransposon and containing a promoterless lacZ reporter gene was constructed to analyse Mycoplasma genitalium transcription in vivo. Recovered transposon insertions were in monocopy, with 16 % expressing enough beta-galactosidase (beta-Gal) to yield colonies exhibiting a detectable blue colour. A sample of 52 blue colonies was propagated and selected for further analyses. The P-Gal activity of the corresponding cultures was measured to quantify, in a reproducible way, the transcription levels of the interrupted ORFs. Several insertions were found in sense with the interrupted ORIF, but surprisingly there was also a number of insertions in non-coding regions, many of them in repetitive DNA regions known as MgPa islands. Moreover, 30% of the analysed transposon insertions had the lacZ gene in the opposite orientation to the coding frame, suggesting the existence of antisense transcripts that may be involved in the control of gene expression in M. genitalium.
